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United States Department of Agriculture

Agricultural Research Service

Title: Assay Methods for Alternanase

Authors
item Ahlgren, Jeffrey
item Cote, Gregory

Submitted to: American Chemical Society National Meeting
Publication Type: Abstract Only
Publication Acceptance Date: August 30, 2001
Publication Date: N/A

Technical Abstract: Alternanase is an endoglucanase that hydrolyzes the alpha-(1-3),(1-6)-D- glucan alternan, giving rise to a series of oligosaccharides that includes the cyclic tetrasaccharide cyclo{-6)-alpha-D-Glcp-(1-3)-alpha-D-Glcp-(1-6)- alpha-D-Glcp-(1-3)-alpha-D-Glcp-(1-}. We recently reported on the ability of alternanase to convert panose to cyclic tetrasaccharide with the concomitant release of D-glucose (G.L. Cote, J. Ahlgren, P. Biely, Abstr. 220th Natl. Meet., ACS, Washington, DC, Aug. 2000, CARB 82). The discovery of this reaction provides us with a specific method for the quantitative analysis of alternanase activity that overcomes the problems inherent in previously used assays. By coupling this reaction with the colorimetric glucose oxidase assay of D-glucose, we are able to obtain an accurate estimate of the specific activity of alternanase preparations. This has not been possible with assays based on reducing values or hydrolysis of dyed alternan. A comparison of the advantages and drawbacks of several assay methods for alternanase will be presented.

Last Modified: 4/18/2014
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